A comparison of Direct sequencing, Pyrosequencing, High resolution melting analysis, TheraScreen DxS, and the K-ras StripAssay for detecting <it>KRAS</it> mutations in non small cell lung carcinomas

• Main Authors: Jancik Sylwia, Drabek Jiri, Berkovcova Jitka, Xu Yong, Stankova Marcela, Klein Jiri, Kolek Vitezslav, Skarda Josef, Tichy Tomas, Grygarkova Ivona, Radzioch Danuta, Hajduch Marian
• Format: Article
• Language: English
• Published: BMC 2012-09-01
• Series: Journal of Experimental & Clinical Cancer Research
• Subjects: SNP - single nucleotide polymorphism; KRAS - Kiras2 kristen rat sarcoma viral oncogene homolog; NSCLC - Non-small cell lung cancer; Genotyping
• Online Access: http://www.jeccr.com/content/31/1/79

<p>Abstract</p> <p>Background</p> <p>It is mandatory to confirm the absence of mutations in the <it>KRAS</it> gene before treating metastatic colorectal cancers with epidermal growth factor receptor inhibitors, and similar regulations are being considered for non-small cell lung carcinomas (NSCLC) and other tumor types. Routine diagnosis of <it>KRAS</it> mutations in NSCLC is challenging because of compromised quantity and quality of biological material. Although there are several methods available for detecting mutations in <it>KRAS</it>, there is little comparative data regarding their analytical performance, economic merits, and workflow parameters.</p> <p>Methods</p> <p>We compared the specificity, sensitivity, cost, and working time of five methods using 131 frozen NSCLC tissue samples. We extracted genomic DNA from the samples and compared the performance of Sanger cycle sequencing, Pyrosequencing, High-resolution melting analysis (HRM), and the Conformité Européenne (CE)-marked TheraScreen DxS and K-ras StripAssay kits.</p> <p>Results and conclusions</p> <p>Our results demonstrate that TheraScreen DxS and the StripAssay, in that order, were most effective at diagnosing mutations in <it>KRAS</it>. However, there were still unsatisfactory disagreements between them for 6.1% of all samples tested. Despite this, our findings are likely to assist molecular biologists in making rational decisions when selecting a reliable, efficient, and cost-effective method for detecting <it>KRAS</it> mutations in heterogeneous clinical tumor samples.</p>